The long-term objective is to develop a national animal resource to provide consistent supply of live and healthy squids for neuroscience and other biomedical research, primarily through the innovative approach of mariculture and secondarily by capture and maintenance of wild squids. Loliginid squids are renowned for their giant fiber system that serves as a fundamental model for study of the nervous systems and the cell membrane. Demand for squids is strong and it is increasing, but supply is very limited. This project is intended to increase the numerical and seasonal availability of live squids, as well as providing the added benefits of supplying research animals of known age, background and good health. The primary specific aim of the renewal is to culture the large loliginid squid Sepioteuthis lessoniana from eggs to adult size and, mainly through laboratory spawning, provide a source of eggs for continuous culture. In a preliminary experiment that represents a breakthrough in cephalopod mariculture, these squids were grown to 1.1 kg in 4-1/2 months, an extraordinary growth rate for any animal. This is 10X the weight of the squid Loligo grown to adult size in 16 months. Furthermore, the giant axons were larger (700 mum), there was no disease and mating and egg laying resulted in more than 3,000 viable embryos. To accomplish this primary aim, experiments and evaluations will be undertaken on: quality and stocking densities of food for hatchlings; refined tank design for hatchlings; microbiological studies of healthy vs. moribund hatchlings; pathogens causing secondary infection in juveniles and adults; prevention and treatment protocols for diseased squids. A major effort will be made to develop a prepared diet for squids to reduce high costs associated with live foods; these experiments will involve behavioral conditioning, chemical attractants and nutrition trials. All research will be conducted in recirculating systems that maintain high-quality sea water. The sepioid squid Sepia officinalis will continue to be cultured through the life cycle and the small squid Lolliguncula brevis will continue to be supplied from local wild stocks. These three species offer a diverse range of attributes for biomedical experimentation, and their production will be increased so that squids of a wide size range will be available on a predictable schedule. In the latter grant period a cost-recovery program will be initiated and the laboratory will operate as a national animal resource center.